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Updated: Apr 19, 2026
Intravital Imaging of Axonal Interactions with Microglia and Macrophages in a Mouse Dorsal Column Crush Injury
Published on: November 23, 2014
Intravital imaging of axonal interactions with microglia and macrophages in a mouse dorsal column crush injury
Teresa A Evans1, Deborah S Barkauskas2, Jay T Myers3
1Department of Neurosciences, Case Western Reserve University.
Abstract:
Traumatic spinal cord injury causes an inflammatory reaction involving blood-derived macrophages and central nervous system (CNS)-resident microglia. Intra-vital two-photon microscopy enables the study of macrophages and microglia in the spinal cord lesion in the living animal. This can be performed in adult animals with a traumatic injury to the dorsal column. Here, we describe methods for distinguishing macrophages from microglia in the CNS using an irradiation bone marrow chimera to obtain animals in which only macrophages or microglia are labeled with a genetically encoded green fluorescent protein. We also describe a injury model that crushes the dorsal column of the spinal cord, thereby producing a simple, easily accessible, rectangular lesion that is easily visualized in an animal through a laminectomy. Furthermore, we will outline procedures to sequentially image the animals at the anatomical site of injury for the study of cellular interactions during the first few days to weeks after injury.
Insights
This study details methods for distinguishing macrophages and microglia in the central nervous system (CNS) after spinal cord injury. These techniques allow for the study of cellular interactions in living animals following traumatic injury.
Area of Science:
- Neuroscience
- Immunology
- Cell Biology
Background:
- Traumatic spinal cord injury triggers an inflammatory response involving blood-derived macrophages and central nervous system (CNS)-resident microglia.
- Understanding the distinct roles of these cell types is crucial for developing effective therapies.
- Intra-vital two-photon microscopy offers a powerful tool for observing these cellular dynamics in real-time within living organisms.
Purpose of the Study:
- To present refined methodologies for differentiating between macrophages and microglia in the CNS following injury.
- To establish a reproducible model for studying cellular interactions post-spinal cord injury.
- To enable longitudinal imaging of the injury site for detailed cellular analysis.
Main Methods:
- Utilizing an irradiation bone marrow chimera model to selectively label either macrophages or microglia with green fluorescent protein.
- Establishing a dorsal column crush injury model in adult animals, creating a well-defined lesion for visualization.
- Implementing intra-vital two-photon microscopy for sequential imaging of the injury site in live animals.
Main Results:
- The described methods allow for clear distinction between blood-derived macrophages and CNS-resident microglia in the injured spinal cord.
- The dorsal column crush model provides a consistent and accessible lesion for microscopic examination.
- The protocol facilitates the observation of cellular interactions over the initial weeks post-injury.
Conclusions:
- This work provides essential tools and models for dissecting the inflammatory response after spinal cord injury.
- The ability to differentiate and track macrophages and microglia is key to understanding their contribution to injury and repair.
- These methods pave the way for future research into targeted therapeutic strategies for spinal cord injury.
